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    EZ-96 DNA Methylation-Lightning Kit

    Highlights

    • Fastest 96-well bisulfite conversion kit for complete, high-throughput (96-well) bisulfite conversion of DNA for methylation analysis.
    • Ready-to-use conversion reagent is added directly to DNA.
    • High-yield, converted DNA is ideal for PCR, Methylation Specific PCR (MSP), arrays, library preps, Next-Generation sequencing, etc.
    Zymo Research's 100% Satisfaction Guarantee

    Original Manufacturer

    Innovated in California, Made in the USA

    Satisfaction 100% guaranteed, read Our Promise

    Highlights

    • Fastest 96-well bisulfite conversion kit for complete, high-throughput (96-well) bisulfite conversion of DNA for methylation analysis.
    • Ready-to-use conversion reagent is added directly to DNA.
    • High-yield, converted DNA is ideal for PCR, Methylation Specific PCR (MSP), arrays, library preps, Next-Generation sequencing, etc.
    Zymo Research's 100% Satisfaction Guarantee

    Original Manufacturer

    Innovated in California, Made in the USA

    Satisfaction 100% guaranteed, read Our Promise

    Cat # Name Size
    D5032 EZ-96 DNA Methylation-Lightning Kit (Shallow-Well) 2 x 96 Rxns.
    D5033 EZ-96 DNA Methylation-Lightning Kit (Deep-Well) 2 x 96 Rxns.
    Cat # Name Size
    D5032-1 Lightning Conversion Reagent 15 ml
    D5031-5 L-Desulphonation Buffer 40 ml
    D5006-3 M-Binding Buffer 125 ml
    D5007-6 M-Elution Buffer 8 ml
    D5007-4 M-Wash Buffer 36 ml
    C2001 Silicon-A Plate 2 Plates
    C2004 Zymo-Spin I-96 Plate 2 Plates
    C2005 Conversion Plate w/ Cover Foil 2 Plates/Foils
    C2002 Collection Plate 2 Plates
    C2003 Elution Plate 2 Plates

    Description

    The EZ-96 DNA Methylation-Lightning Kit is a rapid, high-throughput (96-well) bisulfite conversion kit for DNA methylation analysis. The streamlined workflow features ready-to-use Lightning Conversion Reagent and a combined DNA denaturation and bisulfite conversion process. Desulphonation and clean-up of the converted DNA is performed using a unique 96-well spin-plate. With the Deep-Well Kit (Cat. D5033), samples can be eluted in as little as 15 µl. High yield, converted DNA is ideal for PCR, array, and Next-Generation sequencing, etc.

    Performance

    Technical Specifications
    Applications Purified, converted DNA is of high-quality and well-suited for downstream processes, including library preparation for Next-Generation sequencing, PCR amplification, etc.
    Conversion > 99.5%
    Elution Volume Deep-well: ≥ 15 µl
    Shallow-well: ≥ 30 µl
    Equipment Thermocycler with heated lid, swinging-bucket centrifuge with plate carriers.
    Input 100 pg - 2 µg of DNA.
    Processing Time 1.5 hours
    Recovery > 80%
    Sample Source Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free.

    Resources

    Documents

    FAQ

    ZymoTaq DNA Polymerase has been specifically designed for use in bisulfite amplification reactions. ZymoTaq is available as a stand-alone polymerase (E2001/E2002), PreMix (E2003/E2004), and qPCR PreMix (E2054/E2055).

    Bisulfite conversion will work regardless of context, so the kits are compatible with genomic DNA derived from plants and other species with high non-CpG methylation levels.

    Leaving the desulphonation buffer on the column longer than recommended will cause more degradation and subsequently result in lower yields.

    > 50 bp.

    For best results, keep the method of quantification consistent before and after bisulfite treatment:

    • If quantifying with a NanoDrop, use dsDNA settings (50 μg/ml for Ab260 = 1.0) before treatment and use RNA settings (40 μg/ml for Ab260 = 1.0) after treatment.
    • If quantifying with Qubit, use a dsDNA assay before treatment and use a ssDNA assay after treatment.
    Following bisulfite treatment of DNA, nonmethylated cytosine residues are converted into uracil. The recovered DNA is typically A, U, and T-rich. The original base-pairing no longer exists. Instead, the converted DNA is single stranded with limited non-specific base-pairing at room temperature. Therefore, traditional dsDNA methods of quantification will not accurately represent the amount of recovered bisulfite converted DNA.

    Following bisulfite treatment, DNA will be single stranded with limited non-specific base pairing at room temperature. To visualize, run the converted DNA on an agarose gel then chill the gel on ice or in an ice bath for 30 minutes. This will force enough base-pairing to allow intercalation of the ethidium bromide for the DNA to be visible. If using a Bioanalyzer or TapeStation instrument, use RNA kits and reagents to visualize the converted DNA.

    Poor conversion efficiency and low yields can be due to a variety of different experiment-specific conditions. Please contact Technical Support to discuss your specific experimental conditions and further troubleshoot with a product specialist.

    Converted DNA eluted in M-Elution Buffer can generally be stored at -20°C for 1-3 months. If longer term storage is necessary, we recommend storing at or below -70°C if possible. Bisulfite converted DNA is less stable than dsDNA; for best results, minimize freeze-thawing of converted DNA and use as soon as possible for downstream analysis.

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